Yeast as an alternative expression system

Kevin Pumiglia pumiglk at aa.wl.com
Thu May 4 09:49:45 EST 1995


In article <Pine.OSF.3.91.950502085408.17119A-100000 at leonis.nus.sg>,
mcbsinwc at LEONIS.NUS.SG (Sin Wun Chey) wrote:

> Dear netters,
> I have a big protein to express (aprrox. 150kDa). The E. coli GST 
> expression system is prone to breakdown of large proteins and has the 
> disadvantage of not being post-translationally modified. Have 
> anyone tried the GST-expression system in yeast? It seems to combine the 
> benefits of a quickie job (unlike baculovirus) and the production of 
> a translationally modified protein.
> Any suggestions welcome!
> 
> Thanks,
> Wunchey
>  

We have done this with several proteins and it has worked very nicely (pure
protein, minimal braekdown, cheap and easy to constructn and grow).  The
vector we are using were generously provided by Bert O'Malley's Lab at
Baylor (see Biotechniques, last Feb. maybe?).  One draw back of this
particular vector is that the vectors have no thrombin cleavage site.  The
yield is lower than E. coli, but probably better than Sf9 cell expressed
protein.  Hope this helps! -- Kevin



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